Rapid Detection of Ara h2 Using Single Walled Carbon Nanotube Based Biosensor for Peanut Allergen Control

Abstract

Peanut protein Ara h2 is one of the serious food allergens contributing to the allergic reaction, causing the common reasons for food-related life threating problems over the world. The objective of this study is to develop single-walled carbon nanotubes (SWCNTs) based biosensor using 1-pyrenibutanoic acid succinimidyl ester (1-PBASE) as a linker for rapid detecting peanut allergen Ara h2 in foods. The detection principal of this biosensor was based upon the binding of Ara h2 to the anti-Ara h2 on the linker-modified SWCNTs connecting gold electrode in biosensor silicon template. The measurements of each step were achieved using linear sweep voltammetry (LSV) with a potentiostat for Ara h2 detection. The application of anti-Ara h2 as pAbs on linker immobilized SWCNTs surface increased the sensor resistance values. The developed biosensor showed significant resistance response with the increase in the concentration of Ara h2 from 0 to 1000 ng/mL. Indirect enzyme-linked immunosorbent assay (ELISA) was successfully applied to confirm the specificity of antibodies using 96-well microplates. Scanning electrone microscopy (SEM) confirmed the microstructure of SWCNTs. Sensor sensitivity and specificity were tested with different peanut allergens and the detection range (1-1000 ng/mL).